Formulations of 6-mercaptopurine

ABSTRACT

The present invention provides improved formulations of 6-mercaptopurine that exhibit better bioavailability and faster dissolution than previous formulations.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of provisional application Ser. No.60/558,447, filed Apr. 1, 2004, which is incorporated herein byreference.

FIELD OF THE INVENTION

The present invention relates to a process for preparing improvedformulations of 6-mercaptopurine as well as pharmaceutical compositionscomprising the improved formulations of 6-mercaptopurine where theimproved formulations exhibit a faster release of 6-mercaptopurine underaqueous conditions than prior art formulations and exhibit morefavorable bioavailability profiles than prior art formulations.

BACKGROUND OF THE INVENTION

6-mercaptopurine (6-MP) is a synthetic analogue of natural purine bases.After absorption into the body, it is transformed into nucleotides whichinterfere with nucleic acid biosynthesis, especially in the active Sphase. As such, it used to slow the growth of cancerous cells. 6-MP isindicated as a monotherapy and as part of combination therapies fortreating acute lymphocytic leukemia in both adults and children(Physician's Desk Reference 57^(th) Edition, 2003, page 1615-1618). 6-MPalso exhibits immunosuppressive properties. While it is not officiallyindicated for diseases where treatment with immunosuppressive agents isbeneficial, 6-MP has been widely used for several such conditions,especially for Crohn's disease and colitis.

6-MP is administered orally and has partial and variable absorption andbioavailability. Approximately 50% of an oral dose is absorbed. 6-MP isfurther subject to metabolism, especially by thiopurinemethyltransferase.

The need for improving the therapeutic potential of 6-MP has been knownfor a long time. U.S. Pat. Nos. 4,443,435 and 5,120,740, among others,describe the preparation of prodrugs for 6-MP as ways of improving theuse of this potent drug. Work of this sort continues, as is seen in U.S.Patent Application Publications 20040013728, 20030232760, and20020013287. U.S. Pat. Nos. 6,680,302; 6,576,438; and 6,355,623 describemethods of improving the therapeutic outcome of 6-MP treatment inleukemia and in bowel diseases such as Crohn's disease or colitis bymonitoring metabolites of the 6-MP and/or thiopurine methyltransferaseactivity and setting dosing based on the results. U.S. Pat. Nos.6,692,771 and 6,680,068 and U.S. Patent Application Publications20030077306 and 20020160049 describe emulsion formulations that may helpthe penetration of 6-MP into the body, while U.S. Pat. Nos. 6,602,521and 6,372,254, and U.S. Patent Application Publications 20030133976 and20020164371 describe drug delivery systems that might improve thetherapeutics of 6-MP. None of these latter patents show datademonstrating improved bioavailability or therapeutic outcomes with6-MP. The need still exists for formulations for improved delivery of6-MP that improve the bioavailability thereof.

Standard 6-MP tablets (described in Physician's Desk Reference 57^(th)Edition, 2003, page 1615-1618) reach full dissolution after about anhour under acidic dissolution conditions using a USP type II dissolutionunit with paddles rotating at 50 rpm. 50% dissolution is reached atbetween 10 and 15 minutes. This rate of dissolution is not as fast aswould be desirable. One method of improving the rate of dissolution ofpoorly soluble powders is to micronize them. In the case of 6-MP,micronization does little to improve the rate of dissolution offormulated tablets when compared to the standard formulation. The lackof improved rate of dissolution makes such tablets unlikely to showimproved bioavailability when compared to the standard formulation.Further improvements to the formulation are clearly needed.

SUMMARY OF INVENTION

The present invention is directed to compositions of 6-mercaptopurinewhich give improved rates of dissolution when tested in a dissolutionbath. It has been found that by granulating solutions of6-mercaptopurine and pharmaceutical carriers, and forming tabletstherefrom, compositions are produced that improve the rate ofdissolution of the 6-mercaptopurine. It has been further found thatimprovement in the rate of dissolution of the 6-mercaptopurine leads toan improvement in the bioavailability of the 6-mercaptopurine.

In one embodiment, the invention relates to a pharmaceutical compositioncomprising 6-mercaptopurine wherein the dissolution of the6-mercaptopurine is greater than 50% within seven minutes when thedissolution of a tablet comprising 50 mg of 6-mercaptopurine is measuredin 900 ml of 0.1N HCl at 37° C. in a USP type II device using paddlesrotating at 50 rpm.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the time to reach 50%dissolution of the 6-mercaptopurine is reduced by at least about 30%compared to the standard formulation when the dissolution of a tabletcomprising the pharmaceutical composition comprising 6-mercaptopurine ismeasured in 900 ml of 0.1N HCl at 37° C. in a USP type II device usingpaddles rotating at 50 rpm.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the bioavailability isimproved by at least about 15% when dosed to a mammal as compared to thestandard formulation.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine and a potassium, sodium,magnesium, ammonium, or calcium salt of a pharmaceutically acceptableacid.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine and a potassium, sodium,magnesium, ammonium, or calcium salt of a pharmaceutically acceptableacid wherein the composition exhibits enhanced solubility in aqueousacid as compared to the standard formulation. In one embodiment, thepharmaceutically acceptable acid selected from the group consisting ofacetic acid, ascorbic acid, benzoic acid, citric acid, and tartaricacid.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine and potassium citrate.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder wherein the spray granulation was carried out in afluidized bed.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder wherein the solvent for the solution of 6-mercaptopurinecomprises a solvent selected from the group consisting ofdimethylformamide, dimethylacetamide, dimethylsulfoxide, and mixturesthereof.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder wherein the solvent for the solution of 6-mercaptopurinecomprises a solvent selected from the group consisting of water and anat least about stoichiometric amount of a pharmaceutically acceptablebase, ethanol and an at least about stoichiometric amount of apharmaceutically acceptable base, and ethanol/water mixtures and an atleast about stoichiometric amount of a pharmaceutically acceptable base.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder wherein the solvent for the solution of 6-mercaptopurinecomprises a solvent selected from the group consisting ofethanol/water/potassium hydroxide, ethanol/water/sodium hydroxide, andethanol/potassium hydroxide.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder wherein the solvent for the solution of 6-mercaptopurinecomprises ethanol/potassium hydroxide or ethanol/water/potassiumhydroxide.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder wherein the pharmaceutical carrier powder comprises apowder selected from the group consisting of lactose, starch,microcrystalline cellulose, calcium phosphate, powdered cellulose,sorbitol, and sucrose.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto a pharmaceutical carrier powderthat comprises lactose or microcrystalline cellulose.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder wherein the pharmaceutical carrier powder was pre-sprayedwith a solution of a pharmaceutically acceptable acid in a molar amountthat is greater than the molar amount of potassium hydroxide or otherpharmaceutically acceptable base in the 6-mercaptopurine solutionapplied to the pharmaceutical carrier powder.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder wherein the pharmaceutical carrier powder was pre-sprayedwith a solution comprising an acid selected from the group consisting ofacetic acid, ascorbic acid, benzoic acid, citric acid, and tartaricacid.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising 6-mercaptopurine wherein the 6-mercaptopurine wasspray granulated from a solution onto an acceptable pharmaceuticalcarrier powder wherein the pharmaceutical carrier powder was pre-sprayedwith a solution of citric acid.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising about 3% to about 20% of 6-mercaptopurine andabout 4% to about 30% of potassium citrate.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising about 8% 6-mercaptopurine and about 5% potassiumcitrate.

In another embodiment, the invention relates to a pharmaceuticalcomposition comprising about 3% to about 20% of 6-mercaptopurine whereinthe 6-mercaptopurine was spray granulated from solution onto anacceptable pharmaceutical carrier powder wherein the pharmaceuticalcarrier powder was pre-sprayed with a solution of citric acid.

In another aspect of the invention, the invention relates to a method ofmaking a pharmaceutical composition of 6-mercaptopurine comprising thespray granulation of a solution of 6-mercaptopurine onto apharmaceutical carrier.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine comprising the spraygranulation of a solution of 6-mercaptopurine onto a pharmaceuticalcarrier wherein the 6-mercaptopurine is dissolved in a solvent thatcomprises a solvent selected from the group consisting ofdimethylformamide, dimethylacetamide, dimethylsulfoxide, and mixturesthereof.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine comprising the spraygranulation of a solution of 6-mercaptopurine onto a pharmaceuticalcarrier wherein the 6-mercaptopurine is dissolved in a solvent thatcomprises a solvent selected from the group consisting of water and anat least about stoichiometric amount of a pharmaceutically acceptablebase, ethanol and an at least about stoichiometric amount of apharmaceutically acceptable base, and ethanol/water mixtures and an atleast about stoichiometric amount of a pharmaceutically acceptable base.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine comprising the spraygranulation of a solution of 6-mercaptopurine onto a pharmaceuticalcarrier wherein the 6-mercaptopurine is dissolved in a solvent thatcomprises a solvent selected from the group consisting ofethanol/water/potassium hydroxide, ethanol/water/sodium hydroxide, andethanol/potassium hydroxide. In certain embodiments, the solventconsists essentially of ethanol/water/potassium hydroxide,ethanol/water/sodium hydroxide, or ethanol/potassium hydroxide.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine comprising the spraygranulation of a solution of 6-mercaptopurine onto a pharmaceuticalcarrier wherein the 6-mercaptopurine is dissolved in ethanol/potassiumhydroxide, or ethanol/water/potassium hydroxide.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine comprising the spraygranulation of a solution of 6-mercaptopurine onto a pharmaceuticalcarrier wherein the pharmaceutical carrier comprises a powder selectedfrom the group consisting of lactose, starch, microcrystallinecellulose, calcium phosphate, powdered cellulose, sorbitol, and sucrose.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine comprising the spraygranulation of a solution of 6-mercaptopurine onto a pharmaceuticalcarrier comprising lactose powder or microcrystalline cellulose.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine comprising the spraygranulation of a solution of 6-mercaptopurine onto a pharmaceuticalcarrier wherein the pharmaceutical carrier was pre-sprayed with asolution of a pharmaceutically acceptable acid in a molar amount that isgreater than the molar amount of potassium hydroxide or otherpharmaceutically acceptable base in the 6-mercaptopurine solutionapplied to the pharmaceutical carrier.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine comprising the spraygranulation of a solution of 6-mercaptopurine onto a pharmaceuticalcarrier using a fluidized bed granulator.

In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the 6-mercaptopurine dissolves in 0.1N HCl to anextent of greater than 50% within seven minutes. In another embodiment,the invention relates to a method of spray granulating a solution of6-mercaptopurine onto a pharmaceutical carrier to make a formulation of6-mercaptopurine having enhanced solubility properties such that thetime to reach 50% dissolution of the 6-mercaptopurine formulation isreduced by at least about 30% compared to the standard formulation whenthe dissolution of a tablet comprising 50 mg of 6-mercaptopurine ismeasured in 900 ml of 0.1N HCl at 37° C. in a USP type II device usingpaddles rotating at 50 rpm.

In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the 6-mercaptopurine dissolves in 0.1N HCl to anextent of greater than 50% within seven minutes, wherein the methodcomprises dissolving 6-mercaptopurine in a solvent that comprises asolvent selected from the group consisting of dimethylformamide,dimethylacetamide, dimethylsulfoxide, and mixtures thereof. In certainembodiments, the solvent consists essentially of dimethylformamide,dimethylacetamide, dimethylsulfoxide, or mixtures thereof. In anotherembodiment, the invention relates to a method of spray granulating asolution of 6-mercaptopurine onto a pharmaceutical carrier to make aformulation of 6-mercaptopurine having enhanced solubility propertiessuch that the time to reach 50% dissolution of the 6-mercaptopurineformulation is reduced by at least about 30% compared to the standardformulation when the dissolution of a tablet comprising 50 mg of6-mercaptopurine is measured in 900 ml of 0.1N HCl at 37° C. in a USPtype II device using paddles rotating at 50 rpm, wherein the methodcomprises dissolving 6-mercaptopurine in a solvent that comprises asolvent selected from the group consisting of dimethylformamide,dimethylacetamide, dimethylsulfoxide, and mixtures thereof. In certainembodiments, the solvent consists essentially of dimethylformamide,dimethylacetamide, dimethylsulfoxide, or mixtures thereof.

In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the 6-mercaptopurine dissolves in 0.1N HCl to anextent of greater than 50% within seven minutes, wherein the methodcomprises dissolving 6-mercaptopurine in a solvent that comprises asolvent selected from the group consisting of water and an at leastabout stoichiometric amount of a pharmaceutically acceptable base,ethanol and an at least about stoichiometric amount of apharmaceutically acceptable base, and ethanol/water mixtures and an atleast about stoichiometric amount of a pharmaceutically acceptable base.In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the time to reach 50% dissolution of the6-mercaptopurine formulation is reduced by at least about 30% comparedto the standard formulation when the dissolution of a tablet comprising50 mg of 6-mercaptopurine is measured in 900 ml of 0.1N HCl at 37° C. ina USP type II device using paddles rotating at 50 rpm, wherein themethod comprises dissolving 6-mercaptopurine in a solvent that comprisesa solvent selected from the group consisting of water and an at leastabout stoichiometric amount of a pharmaceutically acceptable base,ethanol and an at least about stoichiometric amount of apharmaceutically acceptable base, and ethanol/water mixtures and an atleast about stoichiometric amount of a pharmaceutically acceptable base.In certain embodiments, the solvent consists essentially of water and anat least about stoichiometric amount of a pharmaceutically acceptablebase, ethanol and an at least about stoichiometric amount of apharmaceutically acceptable base, or ethanol/water mixtures and an atleast about stoichiometric amount of a pharmaceutically acceptable base.

In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the 6-mercaptopurine dissolves in 0.1N HCl to anextent of greater than 50% within seven minutes, wherein the methodcomprises dissolving 6-mercaptopurine in a solvent that comprises asolvent selected from the group consisting of ethanol/water/potassiumhydroxide, ethanol/water/sodium hydroxide, and ethanol/potassiumhydroxide. In another embodiment, the invention relates to a method ofspray granulating a solution of 6-mercaptopurine onto a pharmaceuticalcarrier to make a formulation of 6-mercaptopurine having enhancedsolubility properties such that the time to reach 50% dissolution of the6-mercaptopurine formulation is reduced by at least about 30% comparedto the standard formulation when the dissolution of a tablet comprising50 mg of 6-mercaptopurine is measured in 900 ml of 0.1N HCl at 37° C. ina USP type II device using paddles rotating at 50 rpm, wherein themethod comprises dissolving 6-mercaptopurine in a solvent selected fromthe group consisting of ethanol/water/potassium hydroxide,ethanol/water/sodium hydroxide, and ethanol/potassium hydroxide. Incertain embodiments, the solvent consists essentially ofethanol/water/potassium hydroxide, ethanol/water/sodium hydroxide, orethanol/potassium hydroxide.

In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the 6-mercaptopurine dissolves in 0.1N HCl to anextent of greater than 50% within seven minutes, wherein the methodcomprises dissolving 6-mercaptopurine in ethanol/potassium hydroxide orethanol/water/potassium hydroxide. In another embodiment, the inventionrelates to a method of spray granulating a solution of 6-mercaptopurineonto a pharmaceutical carrier to make a formulation of 6-mercaptopurinehaving enhanced solubility properties such that the time to reach 50%dissolution of the 6-mercaptopurine formulation is reduced by at leastabout 30% compared to the standard formulation when the dissolution of atablet comprising 50 mg of 6-mercaptopurine is measured in 900 ml of0.1N HCl at 37° C. in a USP type II device using paddles rotating at 50rpm, wherein the method comprises dissolving 6-mercaptopurine inethanol/potassium hydroxide or ethanol/water/potassium hydroxide.

In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the 6-mercaptopurine dissolves in 0.1N HCl to anextent of greater than 50% within seven minutes, wherein thepharmaceutical carrier comprises a powder selected from the groupconsisting of lactose, starch, microcrystalline cellulose, calciumphosphate, powdered cellulose, sorbitol and sucrose. In anotherembodiment, the invention relates to a method of spray granulating asolution of 6-mercaptopurine onto a pharmaceutical carrier to make aformulation of 6-mercaptopurine having enhanced solubility propertiessuch that the time to reach 50% dissolution of the 6-mercaptopurineformulation is reduced by at least about 30% compared to the standardformulation when the dissolution of a tablet comprising 50 mg of6-mercaptopurine is measured in 900 ml of 0.1N HCl at 37° C. in a USPtype II device using paddles rotating at 50 rpm, wherein thepharmaceutical carrier comprises a powder selected from the groupconsisting of lactose, starch, microcrystalline cellulose, calciumphosphate, powdered cellulose, sorbitol and sucrose.

In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the 6-mercaptopurine dissolves in 0.1N HCl to anextent of greater than 50% within seven minutes, wherein thepharmaceutical carrier comprises lactose powder. In another embodiment,the invention relates to a method of spray granulating a solution of6-mercaptopurine onto a pharmaceutical carrier to make a formulation of6-mercaptopurine having enhanced solubility properties such that thetime to reach 50% dissolution of the 6-mercaptopurine formulation isreduced by at least about 30% compared to the standard formulation whenthe dissolution of a tablet comprising 50 mg of 6-mercaptopurine ismeasured in 900 ml of 0.1N HCl at 37° C. in a USP type II device usingpaddles rotating at 50 rpm, wherein the pharmaceutical carrier compriseslactose powder or microcrystalline cellulose.

In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the 6-mercaptopurine dissolves in 0.1N HCl to anextent of greater than 50% within seven minutes, wherein thepharmaceutical carrier was pre-sprayed with a solution of apharmaceutically acceptable acid in a molar amount that is greater thanthe molar amount of potassium hydroxide or other pharmaceuticallyacceptable base in the 6-mercaptopurine solution applied to thepharmaceutical carrier. In another embodiment, the invention relates toa method of spray granulating a solution of 6-mercaptopurine onto apharmaceutical carrier to make a formulation of 6-mercaptopurine havingenhanced solubility properties such that the time to reach 50%dissolution of the 6-mercaptopurine formulation is reduced by at leastabout 30% compared to the standard formulation when the dissolution of atablet comprising 50 mg of 6-mercaptopurine is measured in 900 ml of0.1N HCl at 37° C. in a USP type II device using paddles rotating at 50rpm, wherein the pharmaceutical carrier was pre-sprayed with a solutionof a pharmaceutically acceptable acid in a molar amount that is greaterthan the molar amount of potassium hydroxide in the 6-mercaptopurinesolution applied to the pharmaceutical carrier.

In another embodiment, the invention relates to a method of spraygranulating a solution of 6-mercaptopurine onto a pharmaceutical carrierto make a formulation of 6-mercaptopurine having enhanced solubilityproperties such that the 6-mercaptopurine dissolves in 0.1N HCl to anextent of greater than 50% within seven minutes, wherein the spraygranulating uses a fluidized bed granulator. In another embodiment, theinvention relates to a method of spray granulating a solution of6-mercaptopurine onto a pharmaceutical carrier to make a formulation of6-mercaptopurine having enhanced solubility properties such that thetime to reach 50% dissolution of the 6-mercaptopurine formulation isreduced by at least about 30% compared to the standard formulation whenthe dissolution of a tablet comprising 50 mg of 6-mercaptopurine ismeasured in 900 ml of 0.1N HCl at 37° C. in a USP type II device usingpaddles rotating at 50 rpm, wherein the spray granulating uses afluidized bed granulator.

In another aspect of the invention, the invention relates to a method ofmaking a pharmaceutical composition of 6-mercaptopurine having enhancedbioavailability properties such that when dosing said composition to amammal the bioavailability is improved by at least about 15%.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine having enhancedbioavailability properties such that when dosing said composition to amammal the bioavailability is improved by at least about 15%, the methodcomprising the spray granulation of a solution of 6-mercaptopurine ontoa pharmaceutical carrier wherein the 6-mercaptopurine is dissolved in asolvent comprising a solvent selected from the group consisting ofdimethylformamide, dimethylacetamide, dimethylsulfoxide, and mixturesthereof. In certain embodiment, the solvent consists essentially ofdimethylformamide, dimethylacetamide, dimethylsulfoxide, or mixturesthereof.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine having enhancedbioavailability properties such that when dosing said composition to amammal the bioavailability is improved by at least about 15%, the methodcomprising the spray granulation of a solution of 6-mercaptopurine ontoa pharmaceutical carrier wherein the 6-mercaptopurine is dissolved in asolvent comprising a solvent selected from the group consisting of waterand an at least about stoichiometric amount of a pharmaceuticallyacceptable base, ethanol and an at least about stoichiometric amount ofa pharmaceutically acceptable base, and ethanol/water mixtures and an atleast about stoichiometric amount of a pharmaceutically acceptable base.In certain embodiments, the solvent consists essentially of water and anat least about stoichiometric amount of a pharmaceutically acceptablebase, ethanol and an at least about stoichiometric amount of apharmaceutically acceptable base, or ethanol/water mixtures and an atleast about stoichiometric amount of a pharmaceutically acceptable base.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine having enhancedbioavailability properties such that when dosing said composition to amammal the bioavailability is improved by at least about 15%, the methodcomprising the spray granulation of a solution of 6-mercaptopurine ontoa pharmaceutical carrier wherein the solution is 6-mercaptopurinedissolved in a solvent comprising a solvent selected from the groupconsisting of ethanol/water/potassium hydroxide, ethanol/water/sodiumhydroxide, and ethanol/potassium hydroxide.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine having enhancedbioavailability properties such that when dosing said composition to amammal the bioavailability is improved by at least about 15%, the methodcomprising the spray granulation of a solution of 6-mercaptopurine ontoa pharmaceutical carrier wherein the solution is 6-mercaptopurinedissolved in ethanol/potassium hydroxide or ethanol/water/potassiumhydroxide.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine having enhancedbioavailability properties such that when dosing said composition to amammal the bioavailability is improved by at least about 15%, the methodcomprising the spray granulation of a solution of 6-mercaptopurine ontoa pharmaceutical carrier wherein the pharmaceutical carrier comprises apowder selected from the group consisting of lactose, starch,microcrystalline cellulose, calcium phosphate, powdered cellulose,sorbitol, and sucrose.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine having enhancedbioavailability properties such that when dosing said composition to amammal the bioavailability is improved by at least about 15%, comprisingthe spray granulation of a solution of 6-mercaptopurine onto apharmaceutical carrier comprising lactose.

In another embodiment, the invention relates to a method of making apharmaceutical composition of 6-mercaptopurine having enhancedbioavailability properties such that when dosing said composition to amammal the bioavailability is improved by at least about 15%, the methodcomprising the spray granulation of a solution of 6-mercaptopurine ontoa pharmaceutical carrier wherein the pharmaceutical carrier waspre-sprayed with a solution of a pharmaceutically acceptable acid in amolar amount that is greater than the molar amount of potassiumhydroxide or other pharmaceutically acceptable base in the6-mercaptopurine solution applied to the pharmaceutical carrier.

In another embodiment, the invention relates to a method of making apharmaceutical composition comprising 6-mercaptopurine having enhancedbioavailability properties such that when dosing said composition to amammal the bioavailability is improved by at least about 15% compared tothe standard formulation, the method comprising the spray granulation ofa solution of 6-mercaptopurine onto a pharmaceutical carrier using afluidized bed granulator.

In another aspect, the invention relates to a method of dosing apharmaceutical composition comprising 6-mercaptopurine to patients inneed of said drug wherein the composition displays enhanced solubilityin aqueous acid compared to the standard formulation.

In another embodiment, the invention relates to a method of dosing apharmaceutical composition comprising 6-mercaptopurine to patients inneed of said drug wherein the composition displays enhanced solubilityin aqueous acid such that the 6-mercaptopurine dissolves in 0.1N HCl toan extent of greater than 50% within seven minutes or wherein the timeto reach 50% dissolution of the 6-mercaptopurine is reduced by at leastabout 30% compared to the standard formulation when the dissolution of atablet comprising 50 mg of 6-mercaptopurine is measured in 900 ml of0.1N HCl at 37° C. in a USP type II device using paddles rotating at 50rpm.

In another embodiment, the invention relates to a method of dosing apharmaceutical composition comprising 6-mercaptopurine to patients inneed of said drug wherein the bioavailability is improved by at leastabout 15% when dosing to a mammal as compared to the standardformulation.

In another embodiment, the invention relates to a method of dosing apharmaceutical composition comprising 6-mercaptopurine to patients inneed of said drug to treat leukemia or other cancers wherein thecomposition displays enhanced solubility in aqueous acid as compared tothe standard formulation.

In another embodiment, the invention relates to a method of dosing apharmaceutical composition comprising 6-mercaptopurine to patients inneed of said drug to treat Crohn's disease, arthritis, or colitiswherein the composition displays enhanced solubility in aqueous acid ascompared to the standard formulation.

In another embodiment, the invention relates to a method of dosing apharmaceutical composition comprising 6-mercaptopurine to patients inneed of said drug to treat leukemia or other cancers wherein thebioavailability is improved by at least about 15% when dosing to amammal as compared to the standard formulation.

In another embodiment, the invention relates to a method of dosing apharmaceutical composition comprising 6-mercaptopurine to patients inneed of said drug to treat Crohn's disease, arthritis, or colitiswherein the bioavailability is improved by at least about 15% whendosing to a mammal as compared to the standard formulation.

In another embodiment, the invention relates to a method of dosing apharmaceutical composition comprising 6-mercaptopurine to patients inneed of said drug to treat leukemia or other cancers wherein the doseadministered is reduced by at least about 15% and achieves the samebioavailability as the standard formulation.

In another embodiment, the invention relates to a method of dosing apharmaceutical composition comprising 6-mercaptopurine to patients inneed of said drug to treat Crohn's disease, arthritis, or colitiswherein the dose administered is reduced by at least about 15% andachieves the same bioavailability as the standard formulation.

DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the dissolution of a 6-mercaptopurine composition of thepresent invention (6-MP-IB) versus PURINETHOL® in 0.1N HCl (see Example1).

FIG. 2 shows the dissolution of a 6-mercaptopurine composition of thepresent invention from (6-MP-IB batch) vs. PURINETHOL® in 0.1N HCl (seeExample 2).

FIG. 3 shows the average pharmacokinetic profile of 6-mercaptopurine fora pharmaceutical composition of the present invention (6-MP-IB batch)vs. the standard formulation (PURINETHOL®) (see Example 4).

FIG. 4 shows the dissolution of a 6-mercaptopurine tablets prepared asin Example 3. -▴-=PURINETHOL®; -♦-=tablets prepared withmicrocrystalline cellulose; -▪-=tablets prepared with lactose;-x-=lactose tablets, 70% ethanol, 30% water, n=3 (average of threetablets).

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention is directed to compositions of 6-mercaptopurinewhich give improved rate of dissolution when tested in a dissolutionbath and show improved bioavailability characteristics when dosed tomammals.

As used herein, the “standard formulation” is the formulation describedin the Physician's Desk Reference, 57^(th) edition, 2003, pages1615-1618 and sold in the United States under the brand namePURINETHOL®.

As used herein, the term “enhanced solubility properties” or “enhancedsolubility” of a material or composition of the present invention meansan improved rate of dissolution of the material or composition of thepresent invention or an improved extent of dissolution of the materialor composition of the present invention as compared to the standardformulation.

As used herein, the term “improved bioavailability” refers to theincrease in concentration of a drug in the body fluid provided by thecompositions of the present invention as compared to the concentrationof the drug in the body fluid from the standard formulation underidentical conditions. Drug bioavailability is proportional to, and istypically measured by, the total area under the curve (AUC) of theconcentration of the drug found in blood or plasma versus time whenmeasured in a pharmacokinetic trial in a human or an animal. The AUC maybe expressed as AUCt, i.e. the area under the curve to the last measuredtime point, or AUC_(I), i.e. the area under the curve extrapolated toinfinite time. The improvement in bioavailability is measured by thepercent increase in the average AUC of the subjects in the trial whendosing the improved formulation as compared to the average AUC of thesame subjects obtained by dosing of the standard formulation of thedrug. Alternatively, the AUC ratio of the test formulation (AUCf) to theAUC of the reference formulation (AUCr) may be calculated on a persubject basis and then averaged. A percent of the average ratio(AUCf/AUCr) above 100% is then the improvement in bioavailability.

As used herein, the term “slight stoichiometric excess” refers to astoichiometric excess of about 0.1% to about 30%, preferably about 0.5%to about 15%, more preferably about 1% to about 5%, in terms of molepercent.

As used herein, “pre-sprayed” refers to spraying the pharmaceuticalcarrier powder with the acid before the acid-sprayed pharmaceuticalcarrier is contacted with the solution of 6-mercaptopurine.

As used herein, “powder” in reference to a pharmaceutical carrier refersto particles of the pharmaceutical carrier having a size range of 1 to800 microns, more preferably 2 to 500 microns, and most preferably 2 to100 microns or 50 to 400 microns, depending on the material.

One embodiment of the invention is directed to 6-MP formulations thatcomprise 6-MP formulated into granulates by first dissolving the 6-MP inan organic solvent. Examples of solvents that can be used to dissolvethe 6-MP to an extent sufficient to be able to apply the solution to apharmaceutical powder for further processing are dimethylformamide,dimethylacetamide, and dimethylsulfoxide, or mixtures thereof. Lactose,starch, microcrystalline cellulose, calcium phosphate, powderedcellulose, sorbitol or sucrose are examples of pharmaceuticallyacceptable powders that can be used as powders for this granulation.Other pharmaceutical excipient powders are known in the art and may alsobe used. In a more preferred embodiment the organic solvent solution of6-MP is spray granulated on to the powder so as to form a uniformcoating. A preferred method of performing this spray granulation is byusing a fluidized bed granulator. A more preferred embodiment useslactose as the pharmaceutical powder upon which the 6-MP is granulated,and a yet more preferred embodiment uses dimethylformamide to form thegranulation solution. In a more preferred embodiment of the invention alactose granulate is formed that comprises, on a weight/weight (w/w)basis, 1 to 35% 6-MP, more preferably 5 to 20% 6-MP, and most preferablyabout 13% 6-MP. These granulates are then mixed with other tabletexcipients and formed into tablets comprising 0.5 mg to 150 mg of 6-MPfor an approximate tablet weight of 500 mg with an about 50 mg dose themost preferred. Alternately the dose of 6-MP can be controlled bychanging tablet weight using any of the preferred, more preferred, ormost preferred granulates.

Tablets that comprise these formulations of 6-MP have improveddissolution properties. When testing these tablets in 900 ml of 0.1N HClat 37° C. in a USP apparatus II dissolution tester with paddles rotatingat 50 rpm, the rate of dissolution is greatly enhanced compared to thestandard formulation. The time to 50% of dissolution is below sevenminutes more preferably below five minutes and exhibits a more than 30%reduction in the time to 50% dissolution, more preferably a more than50% reduction in time to 50% dissolution, when compared to the standardformulation.

A more preferred embodiment of this invention is directed to 6-MPformulations that comprise 6-MP formulated into granulates by firstdissolving the 6-MP in ethanol containing at least about astoichiometric amount of base, water containing at least about astoichiometric amount of base, or mixtures of ethanol/water containingat least about a stoichiometric amount of base. The base may be selectedfrom any pharmaceutically acceptable base such as the hydroxide orcarbonate salts of potassium, sodium, magnesium, ammonium, or calcium,with potassium hydroxide being preferred. Optionally, a binder such aspolyvinylpyrrolidone (PVP) may be added to the solution. This basicsolution of 6-MP is granulated onto a pharmaceutical carrier selectedfrom the group of lactose, starch, microcrystalline cellulose, calciumphosphate, powdered cellulose, sorbitol, or sucrose. Otherpharmaceutical excipient powders are known in the art and may also beused. In a preferred embodiment, the basic solvent solution of 6-MP isspray granulated on to the powder so as to form a uniform coating. Apreferred method of performing this spray granulation is by using afluidized bed granulator. A more preferred embodiment uses lactose asthe pharmaceutical powder upon which the 6-MP is granulated and a mostpreferred embodiment uses an ethanol/water solvent mixture and potassiumhydroxide as the base. In another more preferred embodiment,microcrystalline cellulose is used as the pharmaceutical powder uponwhich the 6-MP is granulated, and a most preferred embodiment uses anethanol/water solvent mixture and potassium hydroxide as the base. Thebasic granulate is neutralized with a slight stoichiometric excess ofany pharmaceutically acceptable acid. Examples of such acids are aceticacid, ascorbic acid, benzoic acid, citric acid, and tartaric acid. In amore preferred embodiment the acid selected is citric acid. In a morepreferred embodiment, the pharmaceutically acceptable acid is precoatedin a slight stoichiometric excess onto the pharmaceutically acceptablecarrier before it is used in the granulation with the basic organicsolution of 6-MP. In a more preferred embodiment, the pharmaceuticallyacceptable carrier is lactose and the pharmaceutically acceptable acidthat is preloaded in a slight stoichiometric excess is citric acid. Amore preferred mode for applying the acid is spray granulation and amost preferred method uses a fluidized bed granulator. In a preferredembodiment of the invention a lactose granulate is formed that comprises1 to 35% 6-MP, more preferably 5 to 20% 6-MP, and most preferably about11% 6-MP. In another preferred embodiment of the invention, amicrocrystalline cellulose granulate is formed that comprises 1 to 35%6-MP, more preferably 5 to 20% 6-MP, and most preferably about 11% 6-MP.These granulates further comprise salts of pharmaceutically acceptableacids, more preferably the sodium or potassium salts of acetic acid,ascorbic acid, benzoic acid, citric acid, or tartaric acid and mostpreferably the potassium salt of citric acid. The potassium citrate ispresent in about a stoichiometric amount compared to the 6-MP. Thesegranulates are then mixed with other tablet excipients and formed intotablets comprising 0.5 mg to 150 mg of 6-MP for an approximate totaltablet weight of 650 mg with an about 50 mg dose of 6-MP being the mostpreferred. Alternately the dose of 6-MP can be controlled by changingtablet weight using any of the preferred, more preferred, or mostpreferred granulates. In another embodiment the final dosage formcomprises about 3% to about 20% of 6-mercaptopurine and about 2% toabout 30% of potassium citrate and more preferably about 5% to about 15%of 6-MP and about 2% to about 20% potassium citrate, and most preferablyabout 8% 6-mercaptopurine and about 5% potassium citrate.

Tablets that comprise these formulations of 6-MP have improveddissolution properties. When testing these tablets in 900 ml of 0.1N HClat 37° C. in a USP apparatus II dissolution tester with paddles rotatingat 50 rpm, the rate of dissolution is greatly enhanced as compared tothe standard formulation. The time to 50% of dissolution is below sevenminutes, more preferably below five minutes, and exhibits a more than30% reduction in the time to 50% dissolution, more preferably a morethan 50% reduction in time to 50% dissolution, when compared to thestandard formulation.

Another aspect of the invention is a method of producing compositions of6-mercaptopurine which give improved rates of dissolution when tested ina dissolution bath. Standard formulation 6-MP tablets reach fulldissolution after about an hour under acidic dissolution conditionsusing a USP type II dissolution unit with paddles rotating at 50 rpm.50% dissolution is reached at between 10 and 15 minutes Improved ratesof dissolution are defined herein as a time to 50% of dissolution lessthan or equal to about seven minutes, more preferably less than or equalto about five minutes, or a more than 30% reduction in the time to 50%dissolution, more preferably a more than or equal to 50% reduction inthe time to 50% dissolution, compared to the standard formulation.

One aspect of the present invention is a method of forming 6-MPformulations that comprises granulating 6-MP into granulates by firstdissolving the 6-MP in an organic solvent. Examples of solvents that canbe used to dissolve the 6-MP to an extent sufficient to be able to applythe solution to a pharmaceutical powder for further processing aredimethylformamide, dimethylacetamide, and dimethylsulfoxide, or mixturesthereof. Lactose, starch, microcrystalline cellulose, calcium phosphate,powdered cellulose, sorbitol, or sucrose are examples ofpharmaceutically acceptable powders that can be used as powders for thisgranulation. Other pharmaceutical excipient powders are known in the artand may also be used. In a more preferred embodiment the organic solventsolution of 6-MP is spray granulated on to the powder so as to form auniform coating. A preferred method of performing this spray granulationis by using a fluidized bed granulator. A more preferred embodiment useslactose as the pharmaceutical powder upon which the 6-MP is granulatedand a yet more preferred embodiment uses dimethylformamide to form thegranulation solution. In a more preferred embodiment of the invention, alactose granulate is formed that comprises 1 to 35% 6-MP, morepreferably 5 to 20% 6-MP, and most preferably about 13% 6-MP.

These granulates are then mixed with other tablet excipients and formedinto tablets comprising 0.5 mg to 150 mg of 6-MP for an approximatetablet weight of 500 mg with an about 50 mg dose being the mostpreferred. Alternatively, the dose of 6-MP can be controlled by changingtablet weight using any of the preferred, more preferred, or mostpreferred granulates.

Tablets that comprise formulations of 6-MP made by this method haveimproved dissolution properties. When testing these tablets in 900 ml of0.1N HCl at 37° C. in a USP apparatus II dissolution tester with paddlesrotating at 50 rpm, the rate of dissolution is greatly enhanced comparedto the standard formulation. The time to 50% of dissolution is belowseven minutes, more preferably below five minutes, and exhibits a morethan 30% reduction in the time to 50% dissolution, more preferably amore than 50% reduction in time to 50% dissolution, when compared to thestandard formulation.

A more preferred embodiment of this invention is a method of making 6-MPformulations that comprises granulating 6-MP into granulates by firstdissolving the 6-MP in ethanol containing at least a stoichiometricamount of base, water containing at least a stoichiometric amount ofbase, or mixtures of ethanol/water containing at least a stoichiometricamount of base. The base may be selected from any pharmaceuticallyacceptable base such as the hydroxide or carbonate salts of potassium,sodium, magnesium, ammonium, or calcium, with potassium hydroxide beingmore preferred. Optionally, a binder such as polyvinylpyrrolidone (PVP)may be added to the solution. This basic solution of 6-MP is granulatedonto a pharmaceutical carrier selected from the group consisting oflactose, starch, microcrystalline cellulose, calcium phosphate, powderedcellulose, sorbitol, and sucrose. Other pharmaceutical excipient powdersare known in the art and may also be used. In a more preferredembodiment the basic solvent solution of 6-MP is spray granulated on tothe powder so as to form a uniform coating. A preferred method ofperforming this spray granulation is by using a fluidized bedgranulator. A more preferred embodiment uses lactose as thepharmaceutical powder upon which the 6-MP is granulated, and a mostpreferred embodiment uses an ethanol/water solvent mixture and potassiumhydroxide as the base. In another more preferred embodiment,microcrystalline cellulose is used as the pharmaceutical powder uponwhich the 6-MP is granulated, and a most preferred embodiment uses anethanol/water solvent mixture and potassium hydroxide as the base. Thebasic granulate is neutralized with a stoichiometric excess of anypharmaceutically acceptable acid. Examples of such acids are aceticacid, ascorbic acid, benzoic acid, citric acid, and tartaric acid. In amore preferred embodiment, the acid selected is citric acid. In a morepreferred embodiment, the pharmaceutically acceptable acid is preloadedin a slight stoichiometric excess onto the pharmaceutically acceptablecarrier before it is used in the granulation with the basic organicsolution of 6-MP. In a more preferred embodiment, the pharmaceuticallyacceptable carrier is lactose and the pharmaceutically acceptable acidthat is preloaded in a slight stoichiometric excess is citric acid. Amore preferred method for applying the acid is spray granulation, and amost preferred method uses a fluidized bed granulator. In a preferredembodiment of the invention, a lactose granulate is formed thatcomprises 1 to 35% 6-MP, preferably 5 to 20% 6-MP, and most preferablyabout 11% 6-MP. In another preferred embodiment of the invention, amicrocrystalline cellulose granulate is formed that comprises 1 to 35%6-MP, more preferably 5 to 20% 6-MP, and most preferably about 11% 6-MP.These granulates further comprise salts of pharmaceutically acceptableacids, preferably the sodium or potassium salts of acetic acid, ascorbicacid, benzoic acid, citric acid, or tartaric acid, and most preferablythe potassium salt of citric acid. The potassium citrate is present inabout a stoichiometric amount compared to the 6-MP. These granulates arethen mixed with other tablet excipients and formed into tabletscomprising 0.5 mg to 150 mg of 6-MP for an approximate total tabletweight of 650 mg, with an about 50 mg dose of 6-MP in the tablet beingmost preferred. Alternatively, the dose of 6-MP can be controlled bychanging tablet weight using any of the preferred, more preferred, ormost preferred granulates. In another embodiment, the final dosage formcomprises about 3% to about 20% of 6-mercaptopurine and about 2% toabout 30% of potassium citrate, preferably about 5% to about 15% of 6-MPand about 2% to about 20% potassium citrate, and most preferably about8% 6-mercaptopurine and about 5% potassium citrate.

Tablets that comprise formulations of 6-MP made by this method haveimproved dissolution properties. When testing these tablets in 900 ml of0.1N HCl at 37° C. in a USP apparatus II dissolution tester with paddlesrotating at 50 rpm, the rate of dissolution is greatly enhanced comparedto the standard formulation. The time to 50% of dissolution is belowseven minutes, preferably below five minutes, and exhibits a more than30% reduction in the time to 50% dissolution, preferably a more than 50%reduction in time to 50% dissolution, when compared to the standardformulation.

Another aspect of the invention is a method of producing compositions of6-mercaptopurine which provide enhanced bioavailability compared to thestandard formulation. The enhanced bioavailability may be a rise inaverage AUCt or AUC_(I) of about 5% or more, preferably a rise of about15% or more, and most preferably a rise of 20% or more. Alternatively,the average ratio of the individual AUCt values for the test andreference formulations is about 1.05 or more, preferably 1.15 or more,and most preferably 1.20 or more. One embodiment of this aspect of theinvention is a method of making 6-MP formulations that comprisesgranulating 6-MP into granulates by first dissolving the 6-MP in anorganic solvent. Examples of solvents that can be used to dissolve the6-MP to an extent sufficient to be able to apply the solution to apharmaceutical powder for further processing are dimethylformamide,dimethylacetamide, and dimethylsulfoxide, or mixtures thereof. Lactose,starch, microcrystalline cellulose, calcium phosphate, powderedcellulose, sorbitol, or sucrose are examples of pharmaceuticallyacceptable powders that can be used as powders for this granulation.Other pharmaceutical excipient powders are known in the art and may alsobe used. In a more preferred embodiment, the organic solvent solution of6-MP is spray granulated on to the powder so as to form a uniformcoating. A preferred method of performing this spray granulation is byusing a fluidized bed granulator. A more preferred embodiment useslactose as the pharmaceutical powder upon which the 6-MP is granulated,and a yet more preferred embodiment uses dimethylformamide to form thegranulation solution. In a more preferred embodiment of the invention, alactose granulate is formed that comprises 1 to 35% 6-MP, preferably 5to 20% 6-MP, and most preferably about 13% 6-MP. These granulates arethen mixed with other tablet excipients and formed into tabletscomprising 0.5 mg to 150 mg of 6-MP for an approximate total tabletweight of 500 mg, with an about 50 mg of 6-MP in that tablet being thedose most preferred. Alternatively, the dose of 6-MP can be controlledby changing tablet weight using any of the preferred, more preferred, ormost preferred granulates.

A more preferred embodiment of this invention is a method of producing6-MP formulations that comprises granulating 6-MP into granulates byfirst dissolving the 6-MP in ethanol containing at least about astoichiometric amount of base, water containing at least about astoichiometric amount of base, or mixtures of ethanol/water containingat least about a stoichiometric amount of base. The base may be selectedfrom any pharmaceutically acceptable base such as the hydroxide orcarbonate salts of potassium, sodium, magnesium, ammonium, or calcium,with potassium hydroxide being more preferred. Optionally, a binder suchas polyvinylpyrrolidone (PVP) may be added to the solution. This basicsolution of 6-MP is granulated onto a pharmaceutical carrier selectedfrom the group of lactose, starch, microcrystalline cellulose, calciumphosphate, powdered cellulose, sorbitol, and sucrose. Otherpharmaceutical excipient powders are known in the art and may also beused. In a more preferred embodiment the basic solvent solution of 6-MPis spray granulated on to the powder so as to form a uniform coating. Apreferred method of performing this spray granulation is by using afluidized bed granulator. A more preferred embodiment uses lactose asthe pharmaceutical powder upon which the 6-MP is granulated, and a mostpreferred embodiment uses an ethanol/water solvent mixture and potassiumhydroxide as the base. In another more preferred embodiment,microcrystalline cellulose is used as the pharmaceutical powder uponwhich the 6-MP is granulated, and a most preferred embodiment uses anethanol/water solvent mixture and potassium hydroxide as the base. Thebasic granulate is neutralized with a slight stoichiometric excess ofany pharmaceutically acceptable acid. Examples of such acids are aceticacid, ascorbic acid, benzoic acid, citric acid, and tartaric acid. In amore preferred embodiment, the acid selected is citric acid. In a morepreferred embodiment, the pharmaceutically acceptable acid is preloadedin a slight stoichiometric excess onto the pharmaceutically acceptablecarrier before it is used in the granulation with the basic organicsolution of 6-MP. In a more preferred embodiment, the pharmaceuticallyacceptable carrier is lactose and the pharmaceutically acceptable acidthat is preloaded in an about slight stoichiometric excess is citricacid. A more preferred mode for applying the acid is spray granulationand a most preferred method uses a fluidized bed granulator. In apreferred embodiment of the invention a lactose granulate is formed thatcomprises 1 to 35% 6-MP, preferably 5 to 20% 6-MP, and most preferablyabout 11% 6-MP. In another preferred embodiment of the invention, amicrocrystalline cellulose granulate is formed that comprises 1 to 35%6-MP, more preferably 5 to 20% 6-MP, and most preferably about 11% 6-MP.These granulates further comprise salts of pharmaceutically acceptableacids, preferably the sodium or potassium salts of acetic acid, ascorbicacid, benzoic acid, citric acid, or tartaric acid, and most preferablythe potassium salt of citric acid. The potassium citrate is present inabout a stoichiometric amount compared to the 6-MP. These granulates arethen mixed with other tablet excipients and formed into tabletscomprising 0.5 mg to 150 mg of 6-MP for an approximate total tabletweight of 650 mg, with an about 50 mg dose of 6-MP in the tablet beingpreferred. Alternatively, the dose of 6-MP can be controlled by changingtablet weight using any of the preferred, more preferred, or mostpreferred granulates. In another embodiment the final dosage formcomprises about 3% to about 20% of 6-mercaptopurine and about 2% toabout 30% of potassium citrate, preferably about 5% to about 15% of 6-MPand about 2% to about 20% potassium citrate, and most preferably about8% 6-mercaptopurine and about 5% potassium citrate.

Tablets that comprise formulations of 6-MP made by this method haveimproved dissolution properties and improved bioavailability, by morethan 5%, preferably by more than 15%, and most preferably by more than20%, when tested in beagle dogs.

Another aspect of this invention is a method of treating patients inneed of treatment with 6-MP by dosing them with formulations of 6-MPthat have enhanced bioavailability compared to the standard formulation.Examples of patients in need of treatment with 6-MP are patientssuffering from any disease in which a cytotoxic drug is beneficial suchas leukemia, especially acute lymphocytic leukemia, or other cancers, aswell as patients suffering from any disease for which animmunosuppressant drug is beneficial, such as Crohn's diseases,ulcerative colitis, or arthritis.

The enhanced bioavailability may be a rise in average AUCt or AUC_(I) ofabout 5% or more, preferably a rise of about 15% or more, and mostpreferably a rise of about 20% or more. Alternatively, the average ratioof the individual AUCt values for the test and reference formulations isabout 1.05 or more, preferably 1.15 or more, and most preferably about1.20 or more. One embodiment of this aspect of the invention is a methodof dosing, to a mammal, 6-MP formulations that comprise granulates thatwere produced by first dissolving the 6-MP in an organic solvent.Examples of solvents that can be used to dissolve the 6-MP to an extentsufficient to be able to apply the solution to a pharmaceutical powderfor further processing are dimethylformamide, dimethylacetamide, anddimethylsulfoxide, or mixtures thereof. Lactose, starch,microcrystalline cellulose, calcium phosphate, powdered cellulose,sorbitol, or sucrose are examples of pharmaceutically acceptable powdersthat can be used as powders for this granulation. Other pharmaceuticalexcipient powders are known in the art and may also be used. In a morepreferred embodiment the organic solvent solution of 6-MP is spraygranulated on to the powder so as to form a uniform coating. A preferredmethod of performing this spray granulation is by using a fluidized bedgranulator. A more preferred embodiment uses lactose as thepharmaceutical powder upon which the 6-MP is granulated and a yet morepreferred embodiment uses dimethylformamide to form the granulationsolution. In a more preferred embodiment of the invention a lactosegranulate is formed that comprises 1 to 35% 6-MP, more preferably 5 to20% 6-MP and most preferably about 13% 6-MP. These granulates are thenmixed with other tablet excipients and formed into tablets comprising0.5 mg to 150 mg of 6-MP for an approximate tablet weight of 500 mg withan about 50 mg dose the most preferred. Alternately the dose of 6-MP canbe controlled by changing tablet weight using any of the preferred, morepreferred, or most preferred granulates.

Other tablet excipients that may be used to formulate tablets comprisingthe pharmaceutical compositions of the present invention includebinders, diluents, disintegrants, lubricants, colorants, and tastemasking agents. Suitable binders include microcrystalline cellulose,modified celluloses, and povidone. Suitable diluents include calciumhydrogen phosphate (CaHPO₄), anhydrous; lactose; and mannitol. Suitabledisintegrants include sodium starch glycollate (type A), sodium starchglycollate (type B), and crospovidone. Suitable lubricants includesodium stearyl fumarate, dimeticone, macrogol 6000, hydrogenated castoroil, and stearic acid.

A more preferred embodiment of this invention is a method of dosing, toa mammal, 6-MP formulations that comprise granulates that were producedby first dissolving the 6-MP in ethanol containing at least about astoichiometric amount of base, water containing at least about astoichiometric amount of base, or mixtures of ethanol/water containingat least about a stoichiometric amount of base. The base may be selectedfrom any pharmaceutically acceptable base such as the hydroxide orcarbonate salts of potassium, sodium, magnesium, ammonium, or calcium,with potassium hydroxide being preferred. Optionally, a binder such aspolyvinylpyrrolidone (PVP) may be added to the solution. This basicsolution of 6-MP is granulated onto a pharmaceutical carrier selectedfrom the group of lactose, starch, microcrystalline cellulose, calciumphosphate, powdered cellulose, sorbitol and sucrose. Otherpharmaceutical excipient powders are known in the art and may also beused. In a more preferred embodiment the basic solvent solution of 6-MPis spray granulated on to the powder so as to form a uniform coating. Apreferred method of performing this spray granulation is by using afluidized bed granulator. A more preferred embodiment uses lactose asthe pharmaceutical powder upon which the 6-MP is granulated and a mostpreferred embodiment uses an ethanol/water solvent mixture and potassiumhydroxide as the base. In another more preferred embodiment,microcrystalline cellulose is used as the pharmaceutical powder uponwhich the 6-MP is granulated, and a most preferred embodiment uses anethanol/water solvent mixture and potassium hydroxide as the base. Thebasic granulate is neutralized with an about slight stoichiometricexcess of any pharmaceutically acceptable acid. Examples of such acidsare acetic acid, ascorbic acid, benzoic acid, citric acid, and tartaricacid. In a more preferred embodiment, the acid selected is citric acid.In a more preferred embodiment, the pharmaceutically acceptable acid ispreloaded in a slight stoichiometric excess onto the pharmaceuticallyacceptable carrier before it is used in the granulation with the basicorganic solution of 6-MP. In a more preferred embodiment thepharmaceutically acceptable carrier is lactose and the pharmaceuticallyacceptable acid that is preloaded in a slight stoichiometric excess iscitric acid. A more preferred mode for applying the acid is spraygranulation and a most preferred method uses a fluidized bed granulator.In a preferred embodiment of the invention, a lactose granulate isformed that comprises 1 to 35% 6-MP, more preferably 5 to 20% 6-MP, andmost preferably about 11% 6-MP. In another preferred embodiment of theinvention, a microcrystalline cellulose granulate is formed thatcomprises 1 to 35% 6-MP, more preferably 5 to 20% 6-MP, and mostpreferably about 11% 6-MP. These granulates further comprise salts ofpharmaceutically acceptable acids, more preferably the sodium orpotassium salts of acetic acid, ascorbic acid, benzoic acid, citricacid, or tartaric acid and most preferably the potassium salt of citricacid. The potassium citrate is present in about a stoichiometric amountcompared to the 6-MP. These granulates are then mixed with other tabletexcipients and formed into tablets comprising 0.5 mg to 150 mg of 6-MPfor an approximate tablet weight of 650 mg, with an about 50 mg dose themost preferred. Alternatively, the dose of 6-MP can be controlled bychanging tablet weight using any of the preferred, more preferred, ormost preferred granulates. In another embodiment, the final dosage formcomprises about 3% to about 20% of 6-mercaptopurine and about 2% toabout 30% of potassium citrate and more preferably about 5% to about 15%of 6-MP and about 2% to about 20% potassium citrate, and most preferablyabout 8% 6-mercaptopurine and about 5% potassium citrate.

In one embodiment, the patients in need of said treatment are treatedwith a dose similar to the dose given with the standard formulation,thereby achieving enhanced efficacy. In another embodiment, the dose oftreatment is lowered so as to have the same bioavailability as thestandard treatment but achieved with a lower dose of drug. The result ofthe treatment is the same efficacy as the standard formulation with lessexposure to potent drugs and an improved side effect profile.

Methods of making 6-mercaptopurine are known in the art. For example,6-mercaptopurine can be made according to the processes described in G.H. Hitchings, G. B. Elion, U.S. Pat. No. 2,697,702 or G. B. Elion , etal., J. Am. Chem. Soc. 74,411 (1952).

EXAMPLES Example 1 Mercaptopurine Spray Granulated fromDimethylformamide Solution

6-Mercaptopurine (6-MP, Orion-Fermion, 13.2 gm) was dissolved indimethylformamide (DMF, Merck, 1.25 liter) with stirring over a periodof 30 minutes. Lactose (DMV, 85 gm) was charged into a fluidized beddrier/granulator (FBD) and suspended by airflow. The air inlettemperature was 70° C. The DMF solution of 6-MP was sprayed into thesuspended fluidized bed at a rate that maintained a bed temperature of36° C. Total spraying time was 6 hours. The granulated lactose wassubsequently dried in the FBD at 70° C. for one hour and sieved througha 1.0 mm screen. The dry granulate (100 gm which contained 13.2 gm 6-MP)was mixed with potato starch (AVEBE, 25.9 grams), microcrystallinecellulose (Avicel 101, FMC, 13.2 grams) and croscarmellose sodium(Ac-Di-Sol, FMC, 3.7 grams) for 8 minutes. Magnesium stearate (Brenntag,0.5 grams) was added and the powder mixed for a further minute. Thepowder was pressed into tablets using a Korsch 106 rotary tablet press,using 12 mm flat faced round punches with the inscription φβ571. Finaltablet weight was 542 mg and the 6-MP content was 50 mg (6-MP-IB batch131-016-1).

Dissolution analysis was carried out in a USP type II dissolution bath(VanKel) using 900 ml of 0.1N HCl kept at 37° C. and stirred at 50 rpm.Samples were taken at 5, 10 15, 30, 45, and 60 minutes. PURINETHOL®(batch GSKO3C04A) was tested under identical conditions. The 6-MPcontent of the samples was measured by UV spectroscopy at 325 nm againsta standard curve. The results of the measurements are given in Table 1and shown graphically in FIG. 1.

TABLE 1 Dissolution of 6-mercaptopurine from 6-MP-IB 131-016-1 vs.PURINETHOL ® in 0.1N HCl PURINETHOL ® GSK 6-MP-IB 131-016-1 03C04A Time(min) Cumulative % Time (min) Cumulative % 0 0 0 0 5 80 5 27 10 91 10 4815 93 15 59 30 94 30 80 45 94 45 87 60 94 60 92

The results of the dissolution show that the DMF spray granulated 6-MPtablets give a much faster dissolution in 0.1N HCl than the standardformulation tablets. The time to 50% dissolution was better than halvedwith 80% being dissolved in 5 minutes and 91% at 10 minutes. Theimproved speed of dissolution of the product is expected to lead toimproved bioavailability in vivo.

Example 2 Mercaptopurine Spray Granulated from Ethanol/Water/KOHSolution

Citric acid (Merck, 4.6 gm) was dissolved in 69 ml ethanol/water(70:30). This solution was sprayed onto a bed of lactose (DMV, 80 grams)suspended in an FBD granulator using the following conditions: inlet airtemperature 55° C., bed temperature 28° C. 6-mercaptopurine(Orion-Fermion, 11.4 gm) was dissolved in 430 ml ethanol/water (80:20)containing pre-dissolved potassium hydroxide (Merck, 4.0 gram). The 6-MPsolution was then sprayed onto the lactose/citric acid bed in the FBDusing the following conditions: inlet air temperature 55° C., bedtemperature 28° C. The bed was dried in situ at 55° C. for 30 minutes.The dried granulate was passed through a 1.6 mm sieve. The dried andsieved granulate (100 grams) was mixed with potato starch (AVEBE, 26grams), microcrystalline cellulose (Avicel 101, FMC, 11.4 grams),crospovidone (ISP Global Tech, 7.5 grams), and colloidal silicon dioxide(Degussa, 0.5 grams) for 8 minutes. Magnesium stearate (Brenntag, 2.2gram) was added and the powder mixed for a further 2 minutes. The powderwas pressed into tablets using a Korsch 106 rotary tablet press using 12mm flat faced round punches with the inscription φβ571. Final tabletweight was 647 mg and the 6-MP content was 50 mg (6-MP-IB batch131-018-6)

Dissolution analysis was carried out in a USP type II dissolution bath(VanKel) using 900 ml of 0.1N HCl kept at 37° C. and stirred at 50 rpm.Samples were taken at 5, 10, 15, 30, 45, and 60 minutes. PURINETHOL®(batch GSKO3CD4A) was tested under identical conditions. The 6-MPcontent of the samples was measured by UV spectroscopy at 325 nm againsta standard curve. The results of the measurements are given in Table 2and shown graphically in FIG. 2.

TABLE 2 Dissolution of 6-mercaptopurine from 6-MP-IB 131-018-6 vs.PURINETHOL ® in 0.1N HCl PURINETHOL ® GSK 6-MP-IB 131-018-6 03C04A Time(min) Cumulative % Time (min) Cumulative % 0 0 0 0 5 67 5 27 10 91 10 4815 96 15 59 30 98 30 80 45 98 45 87 60 96 60 92

The results of the dissolution show that the basic ethanolic-water spraygranulated 6-MP tablets give a much faster dissolution in 0.1N HCl thanthe standard formulation tablets. The time to 50% dissolution was betterthan halved with 67% being dissolved in 5 minutes and better than 90% at10 minutes. The improved speed of dissolution of the product is expectedto lead to improved bioavailability in vivo.

Example 3 Tablets of 6-MP Coated on Microcrystalline Cellulose orLactose

This example present data from tablets in which 6-MP is coated on eithermicrocrystalline cellulose or lactose. Table 3 shows a batch formula fortablets having 40 mg of 6-MP per tablet (the batch is for ˜1000tablets), tablet weight 523 mg using 50% ethanol by volume (44.4% byweight) in both spraying steps.

TABLE 3 Raw material (g) (g) 1 Lactose monohydrate 280 — 2Microcrystalline Cellulose — 280 3 Citric Acid anhydrate  19.5  19.5 4Alcohol denatured or USP  96^(#)  96^(#) 5 Purified Water 120 120 6Mercaptopurine  40.0  40.0 7 Potassium hydroxide  16.2  16.2 8 PVP K30 — 10.4 9 Alcohol denatured or USP 600^(#) 600^(#) 10 Purified Water 750750 11 Colloidal Silicon Dioxide  1.6  1.6 12 Potato Starch  24.4  24.413 Crospovidone  26.4  26.4 14 Microcrystalline Cellulose  91.6  91.6 15PVP K30  15.6  5.2 16 Magnesium Stearate  8.0  8.0 ^(#)Density 0.8 g/mL

Manufacturing Method Solution A.

Mix alcohol (denatured or USP) (4) with purified water (5), add anddissolve citric acid (3).

Coating step I (Aeromatic Strea 1)

Spray solution A on to lactose monohydrate (1) or microcrystallinecellulose (MCC) (2).

Process parameters:

Atomizing air: 1 bar Nozzle: 1.0 mm

Inlet temperature: 55° C.Exhaust temperature: approx. 24° C.Spray rate: approx. 9-10 g/minAirflow rate: approx. 54 m³/h

Solution B.

Mix alcohol (denatured or USP) (9) with purified water (10), add anddissolve potassium hydroxide (7). Add and dissolve 6-mercaptopurine (6).Optionally, PVP K30 (8) may be dissolved in this solution (either withlactose or with MCC-shown here with MCC).

Coating step II (Aeromatic Strea 1)

Spray solution B onto the lactose monohydrate with citric acid or MCCwith citric acid of coating step I.

Process parameters:

Atomizing air: 1 bar Nozzle: 1.0 mm

Inlet temperature: 55° C.Exhaust temperature: approx. 24° C.Spray rate: approx. 10-11 g/minAirflow rate: approx. 54-80 m³/h

Drying

Dry the lactose/citric acid/potassium hydroxide/6-mercaptopurine mixtureor the MCC/citric acid/potassium hydroxide/PVP/6-mercaptopurine mixture.

Process parameters:Inlet temperature: 55° C.Exhaust temperature: approx. 34° C.Airflow rate: approx. 54-80 m³/h

Sieving I

Pass the lactose/citric acid/potassium hydroxide/6-mercaptopurinemixture or the MCC/citric acid/potassium hydroxide/PVP/6-mercaptopurinemixture through a 1.0 mm sieve.

Pass colloidal silicon dioxide (11) through a 1.0 mm sieve.

Mixing I

Blend the lactose/citric acid/potassium hydroxide/6-mercaptopurinemixture or the MCC/citric acid/potassium hydroxide/PVP/6-mercaptopurinemixture with colloidal silicon dioxide for 2 minutes in a cubic tumbler.

Sieving II

Pass potato starch (12), crospovidone (13), microcrystalline cellulose(14) and PVP K30 (15) through 1.0 mm sieve.

Mixing II

Blend the lactose/citric acid/potassiumhydroxide/6-mercaptopurine/colloidal silicon dioxide mixture or theMCC/citric acid/potassium hydroxide/PVP/6-mercaptopurine/colloidalsilicon dioxide mixture with potato starch, crospovidone,microcrystalline cellulose and PVP K30 for 8 minutes in a cubic tumbler.

Sieving III

Pass magnesium stearate (16) through a 1.0 mm sieve.

Mixing III

Blend the mixture of Mixing step II with magnesium stearate for 2minutes in a cubic tumbler.

Tabletting

Compress the final mixture into tablets with tablet weight 523 mg (12mm, round convex R=9.5). Resistance to crushing of 5-7 Kp, friabilitymax. 1.0%, disintegration time<5 min.

The results of the dissolution of 6-MP tablets prepared as in thisexample in 900 ml 0.1N HCl at 37° C. and 50 rpm is shown in FIG. 4.

Example 4 A Comparative Bioavailability Study of a New Oral Formulationof 6-Mercaptopurine (6-MP-IB) vs. PURINETHOL® in Beagle Dogs

Study Objective—To determine the pharmacokinetic profile (AUCt andAUC_(I), Cmax, Tmax, and half life of 6-mercaptopurine in the plasmafollowing oral ingestion of each formulation to show improvedbioavailability for 6-MP-IBStudy Design—Single center, single dose, non-randomized, open label(blinded to analyst), two treatment, two period crossover comparativebioavailability study.Subjects—Six female beagle dogs, 2-3 years old, 9-11 kg body weight.

Study Administrations

1) PURINETHOL® (GSK): Half of a 50 mg tablet (i.e. 25 mg) of6-mercaptopurine, Lot #A067350.2) 6-MP-IB batch 131-018-6: Half of a 50 mg tablet (i.e. 25 mg) of6-mercaptopurine.

The dogs received the half tablets in the fasted state (twelve hoursfast). The tablets were placed in the back of the dog's throat. About 10ml of water was squirted into the mouth with a syringe to facilitateswallowing. The mouth was examined to ensure that the tablet wasswallowed.

Blood Collection and Handling

Blood samples were taken from an indwelling catheter inserted in thejugular vein at 0 hour and at 0.25, 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 5.0,and 6.0 hours post dosing. Seven milliliters of blood was collected ateach time point. The blood was chilled in ice immediately aftercollection. Within two minutes of collection the blood was transferredto tubes containing EDTA. The blood was processed to obtain the plasmawithin one hour. The plasma was stabilized with dithiothreitol andfrozen to −80° C.

Analyses

The analysis of 6-MP in the plasma was carried out at AnapharmLaboratories by a validated LC/MS/MS method.

Study Duration

Two study sessions with a wash out of two weeks between study sessions.

Results

The results of the analysis of 6-MP in the plasma for all the dogs aregiven in Table 4A for the reference PURINETHOL® and in Table 4B for thetest formulation 6-MP-IB.

The results of the calculated pharmacokinetic parameters from theconcentration data are collected in Table 5 while the results of a perdog ratio analysis are given in Table 6. The average pharmacokineticprofiles for all six dogs for each treatment are given in FIG. 3.

One can see in Table 5 that the average AUCt and AUC_(I) are both about20% higher for the test formulation (i.e., the composition of thepresent invention) when compared to the standard formulation. The Cmaxis almost 70% higher. In the ratio analysis, shown in Table 6, whereeach dog is its own control, there is an average ratio of 1.26 or a 26%rise in the bioavailability of the test versus the reference product.

FIG. 3 shows that the advantage of the faster dissolving formulation inbioavailability is in the early time points with higher drugconcentrations being found shortly after drug ingestion. The T max forthe averaged data is shorter for the test compared to reference despitethe fact that the average Tmax (averaged over the individual dogs) isthe same for the two formulations.

Conclusions

The formulation provided by the present invention has been shown to givea more than 20% increase in bioavailability of 6-mercaptopurine in vivowhen compared to an equivalent dose of the standard formulation. Theimproved bioavailability is expected to allow improved therapeuticoutcomes.

TABLE 4a 6-mercaptopurine standard formulation (PURINETHOL ®)concentrations (ng/ml) Subject Period Draw Times (Hour) # # 0.000 0.2500.500 1.00 1.50 2.00 3.00 4.00 5.00 6.00 02 1 <2.00 35.15 38.98 149.72131.27 80.36 26.90 11.01 7.87 5.37 03 1 <2.00 <2.00 53.24 41.64 31.9639.83 19.10 8.85 4.76 2.73 04 1 <2.00 21.69 112.90 54.94 26.45 15.249.75 12.12 8.24 <2.00 05 1 <2.00 20.97 <2.00 123.11 75.23 62.88 41.1913.16 8.96 4.87 06 1 <2.00 61.09 143.83 106.22 42.88 22.53 8.98 5.843.23 2.19 11 1 <2.00 <2.00 <2.00 59.72 91.79 39.99 10.20 4.53 2.46 2.03

TABLE 4b 6-mercaptopurine (6-MP-IB 131-018-6) concentrations (ng/ml)Subject Period Draw Times (Hour) # # 0.000 0.250 0.500 1.00 1.50 2.003.00 4.00 5.00 6.00 02 2 <2.00 25.07 109.97 181.60 77.10 37.32 15.228.52 5.29 3.83 03 2 <2.00 129.92 159.49 79.27 77.05 37.12 11.66 6.643.62 <2.00 04 2 <2.00 30.68 173.75 99.24 35.45 21.17 8.88 4.35 2.71 8.2905 2 <2.00 <2.00 380.69 172.31 59.78 27.99 20.85 12.50 8.26 5.91 06 2<2.00 <2.00 4.61 104.99 44.09 56.45 19.34 10.30 6.69 4.05 11 2 <2.0070.75 139.59 69.21 24.87 21.03 5.47 3.15 2.14 <2.00

TABLE 5 Pharmokinetic results of dog study of 6-Mercaptopurine Dog-session- AUCt AUCi t½ Tmax Cmax treatment (h * ng/g) (h * ng/g) (h) (h)(ng/g) 02-2-test 235.8 241.7 1.1 1.0 181.6 03-2-test 220.2 220.2 0.9 0.5159.5 04-2-test 176.1 188.2 1.0 0.5 173.8 05-2-test 324.4 338.5 1.7 1.0380.7 06-2-test 154.7 160.6 1.0 1.0 105.0 11-2-test 143.6 143.6 0.9 0.5139.6 02-1-ref 272.6 279.5 0.9 1.0 149.7 03-1-ref 120.7 124.5 1.0 0.553.2 04-1-ref 130.0 130.0 1.7 0.5 112.9 05-1-ref 217.3 224.3 1.0 1.0123.1 06-1-ref 179.8 183.3 1.1 0.5 143.8 11-1-ref 124.0 126.2 0.8 1.591.8 AVG(test) 209.1 215.5 1.1 0.8 190.0 AVG (ref) 174.1 178.0 1.1 0.8112.4

TABLE 6 Ratio Analysis AUCt- Cmaxtest/ test/ Dog Cmaxref AUCt-ref 021.21 0.86 03 3.00 1.82 04 1.54 1.35 05 3.09 1.49 06 0.73 0.86 11 1.521.16 AVG 1.848 1.259

1-58. (canceled)
 59. A method of manufacturing a pharmaceuticalcomposition comprising: (a) combining an aqueous solution of apharmaceutically acceptable acid with a solid carrier; (b) combining thesolid carrier of step (a) with an aqueous solution comprising6-mercaptopurine and a pharmaceutically acceptable base.
 60. The methodof claim 59 wherein step (a) comprises spraying the aqueous solution ofthe pharmaceutically acceptable acid onto the solid carrier.
 61. Themethod of claim 59 wherein step (b) comprises spraying the aqueoussolution comprising 6-mercaptopurine and the pharmaceutically acceptablebase onto the solid carrier of step (a).
 62. The method of claim 59wherein step (a) comprises spraying the aqueous solution of thepharmaceutically acceptable acid onto the solid carrier and step (b)comprises spraying the aqueous solution comprising 6-mercaptopurine andthe pharmaceutically acceptable base onto the solid carrier of step (a).63. The method of claim 59 wherein the pharmaceutically acceptable acidis selected from the group consisting of: acetic acid, ascorbic acid,citric acid and tartaric acid.
 64. The method of claim 59 wherein thepharmaceutically acceptable acid is citric acid.
 65. The method of claim59 wherein the solid carrier is selected from the group consisting of:lactose, starch, microcrystalline cellulose, calcium phosphate, powderedcellulose, sorbitol, and sucrose.
 66. The method of claim 59 wherein thesolid carrier is lactose or microcrystalline cellulose.
 67. The methodof claim 59 wherein the solid carrier is lactose.
 68. The method ofclaim 59 wherein the solid carrier is microcrystalline cellulose. 69.The method of claim 59 wherein the pharmaceutically acceptable base isselected from the group consisting of: hydroxide or carbonate salts ofpotassium, sodium, magnesium, ammonium, or calcium.
 70. The method ofclaim 59 wherein the pharmaceutically acceptable base is potassiumhydroxide.
 71. The method of claim 59 wherein: the pharmaceuticallyacceptable acid is citric acid; the solid carrier is lactose ormicrocrystalline cellulose; and the pharmaceutically acceptable base ispotassium hydroxide.
 72. The method of claim 71 wherein the solidcarrier is lactose.
 73. The method of claim 71 wherein the solid carrieris microcrystalline cellulose.
 74. The method of claim 62 wherein: thepharmaceutically acceptable acid is citric acid; the solid carrier islactose or microcrystalline cellulose; and the pharmaceuticallyacceptable base is potassium hydroxide.
 75. The method of claim 74wherein the solid carrier is lactose.
 76. The method of claim 74 whereinthe solid carrier is microcrystalline cellulose.
 77. The method of claim62 wherein the 6-mercaptopurine forms a coating on the solid carrier.78. The method of claim 62 wherein the 6-mercaptopurine forms a uniformcoating on the solid carrier.